Zhou Xianwu, Chen Zerui, Zhou Jianrong, Liu Yaorong, Fan Ruixin, Sun Tucheng
Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong Provincial Key Laboratory of South China Structural Heart Disease, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.
Department of Cardiovascular Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China.
Front Cardiovasc Med. 2021 May 28;8:627380. doi: 10.3389/fcvm.2021.627380. eCollection 2021.
To investigate the N6-methyladenosine (m6A) modification and the expressions of the m6A regulatory genes in the acute aortic dissection (AD). MeRIP-seq and RNA-seq experiments of aortic media tissue samples obtained from AD ( = 4) and Controls ( = 4) were conducted. m6A methylation quantification was used to measure the total mRNA m6A level. The five m6A regulators mRNA expressions were analyzed by quantitative polymerase chain reaction (qPCR). Western blot analyses and immunofluorescence staining were used to detect the difference of METTL14 protein expression in the aortas of AD and Normal. Among AD patients, we detected significantly elevated levels of m6A in total RNA. Compared with the normal group, the up methylated coding genes of AD were primarily enriched in the processes associated with extracellular fibril organization, while the genes with down methylation were enriched in the processes associated with cell death regulation. Furthermore, many differentially methylated m6A sites (DMMSs) coding proteins were mainly annotated during the extracellular matrix and inflammatory responses. These findings indicate that differential m6A methylation and m6A regulatory genes, including MTEEL14 and FTO, may act on functional genes through RNA modification, thereby regulating the pathogenesis of aortic dissection.
研究急性主动脉夹层（AD）中N6-甲基腺苷（m6A）修饰及m6A调控基因的表达。对从AD患者（n = 4）和对照组（n = 4）获取的主动脉中膜组织样本进行了甲基化RNA免疫沉淀测序（MeRIP-seq）和RNA测序（RNA-seq）实验。采用m6A甲基化定量法测定总mRNA的m6A水平。通过定量聚合酶链反应（qPCR）分析5种m6A调控因子的mRNA表达。采用蛋白质免疫印迹分析和免疫荧光染色检测AD患者与正常人群主动脉中METTL14蛋白表达的差异。在AD患者中，我们检测到总RNA中m6A水平显著升高。与正常组相比，AD患者中甲基化上调的编码基因主要富集在与细胞外纤维组织相关的过程中，而甲基化下调的基因则富集在与细胞死亡调控相关的过程中。此外，许多差异甲基化的m6A位点（DMMSs）编码的蛋白质主要在细胞外基质和炎症反应过程中发挥作用。这些发现表明，包括METTL14和FTO在内的差异m6A甲基化和m6A调控基因可能通过RNA修饰作用于功能基因，从而调控主动脉夹层的发病机制。
